磷酸肌酸钠对慢病毒介导Calumenin蛋白沉默阿霉素损伤心肌细胞内质网应激信号通路的作用  

The effect of sodium phosphocreatine on the endoplasmic reticulum stress signal pathway of adriamycin-injured cardiomyocytes that Calumenin silencing by shRNA

作　　者：王伊林[1] 赵雅君[1] 杨洋[1] 柴花 黄侠[1] 赵明[1] 

机构地区：[1]内蒙古民族大学附属医院心内科,内蒙古通辽028000

出　　处：《临床心血管病杂志》2015年第10期1119-1122,共4页Journal of Clinical Cardiology

摘　　要：目的:探讨磷酸肌酸钠对网腔钙结合蛋白(Calumenin)沉默阿霉素损伤心肌细胞内质网应激信号通路作用。方法:培养乳鼠心肌细胞,构建稳定的慢病毒——Calumenin质粒,转染乳鼠培养心肌细胞,实验分为4组:对照组(正常细胞+3mg/L阿霉素)、模型组(慢病毒感染细胞+3mg/L阿霉素)、磷酸肌酸钠1组(正常细胞+阿霉素+磷酸肌酸钠)、磷酸肌酸钠2组(转染细胞+阿霉素+磷酸肌酸钠)。采用Western blotting技术检测各组心肌细胞Calumenin蛋白、内质网应激伴侣蛋白GRP78及内质网应激信号通路因子PERK、PATF-4PERK、eIF2ɑ、ATF-4、IRE1、CHOP表达。结果:1与对照组比较,模型组及磷酸肌酸钠2组心肌细胞Calumenin蛋白表达明显减少(P<0.01)。2与对照组相比,模型组内质网应激伴侣蛋白GRP78及内质网应激信号通路因子PPERK、eIF2ɑ、ATF-4、IRE1、CHOP表达明显增多(P<0.01)。3与模型组相比较,磷酸肌酸钠1组及磷酸肌酸钠2组内质网应激伴侣蛋白GRP78及内质网应激信号通路因子P-PERK、eIF2ɑ、ATF-4、IRE1、CHOP表达减少(P<0.01)。结论:阿霉素损伤可能诱发ERS,并通过ERS凋亡信号通路PERK→P-PERK→eIF2a→ATF-4→CHOP/IRE1→CHOP引起心肌细胞凋亡;磷酸肌酸钠可抑制阿霉素损伤所诱导内质网应激介导的心肌细胞凋亡,这一作用机制可能是通过Calumenin蛋白抑制ERS及其凋亡信号通路PERK→P-PERK→eIF2a→ATF-4→CHOP/IRE1→CHOP实现的。Objective：To investigate the effect of sodium phosphocreatine on the endoplasmic reticulum stress signal pathway of adriamycin-injured cardiomyocytes that calumenin silencing by shRNA.Method：Firstly,constructing stable lentiviral vectors（calumenin plasmid）,in vitro cultured neonatal rat cardiac myocytes were randomly divided into control group（3mg/L ADR）,lentivirus infection group（lentivirus infection＋3mg/L ADR）,sodium phosphocreatine group 1（3 mg/L ADR＋sodium phosphocreatine）,sodium phosphocreatine group 2（lentivirus infection＋3mg/L ADR＋sodium phosphocreatine）,The expression of calumenin,Endoplasmic reticulum stress chaperone GRP78 and endoplasmic reticulum stress signal pathway PERK、P-PERK 、eIF2ɑ、ATF-4、IRE1、CHOP of ADR-injured cardiomyocytes was monitored by Western blotting.Result：（1）Compared with control group,the expression of calumenin of lentivirus infection group and sodium phosphocreatine group 2was significantly reduced（P〈0.01）.（2）Compared with control group,the expression of endoplasmic reticulum stress chaperone GRP78 and endoplasmic reticulum stress signal pathway P-PERK 、eIF2ɑ、ATF-4、IRE1、CHOP of lentivirus infection group and sodium phosphocreatine group 2 was significantly increased（P〈0.01）.（3）Compared with lentivirus infection group,the expression of Endoplasmic reticulum stress signal pathway P-PERK 、eIF2ɑ、ATF-4、IRE1、CHOP of ADR-injured cardiomyocytes in sodium phosphocreatine group 1and sodium phosphocreatine group 2was reduced（P〈0.01）.Conclusion：Calumenin has a role in the alleviation of ER stress induced by ADRinjured myocardial cells;the sodium phosphocreatine can restrain Endoplasmic reticulum stress caused by adriamycin-injured cardiomyocytes,that may be achieved through the Calumenin protein and endoplasmic reticulum stresssignal pathway of PERK→P-PERK→eIF2a→ATF-4→CHOP/IRE1→CHOP.

关 键 词：磷酸肌酸钠 阿霉素 内质网应激 网腔钙结合蛋白 

分 类 号：R331.3[医药卫生—人体生理学] R-332[医药卫生—基础医学]